LOCUS       Exported                7714 bp DNA     circular SYN 17-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7714)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7714
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    54..63
                     /label=CS2 insertion
                     /note="CS2 insertion"
     misc_feature    64..78
                     /label=B-globin 5' mRNA sequence
                     /note="B-globin 5' mRNA sequence"
     CDS             88..804
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMALL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(134..154)
                     /label=EGFP-N primer
     CDS             811..1527
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(857..877)
                     /label=EGFP-N primer
     primer_bind     1464..1485
                     /label=EGFP-C-FOR primer
     CDS             1534..2250
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1580..1600)
                     /label=EGFP-N primer
     primer_bind     2187..2208
                     /label=EGFP-C-FOR primer
     CDS             2296..2316
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2329..2373
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2398..3702
                     /codon_start=1
                     /label=mMeikin
                     /translation="MDKIWHMGPRGDYTRKKRAGERLNLTPKPDLALPGRTEALPGLKG
                     KGKEQGLRKITEKKELSRLTGSSSQRPSLLSVTGGEHLQENSPGQETPEEKTPPCETVT
                     DTFEMDSLLSSTELVSGPAEQDDFSSCLPSCSNAELHTESTDERGSSFPSPELFRGSDC
                     LDWEHPKLEDYMFYKNSTLLDTSKAVVIEKAPQFANLSAVLSSSSKNYEKRHRKIGMTL
                     AAQHLSPEPKYASNLASVVDNAASEVVFAEKTGPPTTEKTQKKPENESEDSGPLVQTKL
                     SSGHPDNKALCSPLSSALESTAVRYTLLPQPLEPVLKKGCILPDKQSKALLTSTPSSDI
                     AEFVIDLSPVQNVSFEELFPNVSNYVNSSEVVPVSSLQESSSNEFSPNTSEICCIIRSS
                     PGTRQMRRKDPAVKNRCSPPKDVPLDIIMKTNGRT"
     promoter        complement(3741..3758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3763..3897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3841..3860
                     /label=SV40pA-primer-Reverse
     misc_feature    3959..3991
                     /label=donwstream polylinker (from pVZ1)
                     /note="donwstream polylinker (from pVZ1)"
     promoter        complement(4010..4028)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4049..4065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4073..4089
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4097..4127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4142..4163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4451..5039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5210..6070)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6071..6175)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6201..6656)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6730..7714
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccatg gtgagcaagg gcgaggagct gttcaccggg
      121 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      181 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      241 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      301 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      361 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      421 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      481 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
      541 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
      601 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      661 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
      721 cccaacgaga agcgcgatca catggccctg ctggagttcg tgaccgccgc cgggatcact
      781 ctcggcatgg acgagctgta caagacgcgt atggtgagca agggcgagga gctgttcacc
      841 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      901 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
      961 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1021 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1081 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1141 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1201 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1261 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1321 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1381 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     1441 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1501 actctcggca tggacgagct gtacaagccc gggatggtga gcaagggcga ggagctgttc
     1561 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1621 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1681 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1741 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1801 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1861 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1921 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1981 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2041 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2101 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2161 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2221 atcactctcg gcatggacga gctgtacaag tccggagggg atcggtggag ctccaccggt
     2281 ggcggccgct ctagagagaa tctttatttt cagggcgccg ccaaattcaa agaaaccgct
     2341 gctgctaaat tcgaacgcca gcacatggac agcggaggtg gaggtactag tctcgagatg
     2401 gataagatct ggcacatggg acccaggggc gactacacca gaaaaaaaag ggccggcgag
     2461 aggctgaacc tgacccctaa acccgacctg gccctgcctg gaagaaccga agccctgccc
     2521 ggcctcaagg gaaagggcaa ggagcaaggc ctgaggaaga ttacagagaa gaaggagctc
     2581 agcaggctga ccggcagcag ctcccagaga ccttccctgc tcagcgtgac cggaggcgag
     2641 cacctgcagg agaacagccc cggacaggag acccccgagg aaaagacccc cccctgcgag
     2701 accgtcaccg ataccttcga gatggactcc ctgctgtcct ccaccgagct ggtgagcgga
     2761 cccgctgagc aggatgactt ctcctcctgc ctgcccagct gtagcaacgc cgagctgcat
     2821 acagaatcca cagatgagag gggcagcagc ttccccagcc ccgagctgtt caggggcagc
     2881 gactgcctgg actgggagca tcccaagtta gaagactaca tgttctacaa aaattctacc
     2941 ctgctcgata cctccaaagc cgtggtcatc gagaaggccc cccaattcgc caatctgagc
     3001 gccgtcctct ccagctccag caagaactac gagaagagac acagaaagat tggcatgaca
     3061 ctggccgccc agcacctgag ccctgagcct aagtacgcta gcaacctggc ctccgtcgtg
     3121 gataacgccg ccagcgaagt ggtcttcgcc gagaagacag gcccccccac caccgagaag
     3181 acccagaaaa agcccgagaa tgagtccgag gacagcggcc ctctggtgca gaccaagctc
     3241 tccagcggcc atcccgacaa caaggccctg tgtagccctc tgtcctccgc cctggagtcc
     3301 accgctgtga gatacaccct gctgccccag cccctcgaac ctgtgctgaa gaagggctgc
     3361 atcctgcccg acaagcagag caaggctctg ctgacatcca caccctcctc cgatatcgcc
     3421 gagttcgtga tcgacctctc ccctgtccag aacgtgagct ttgaggagct gttccccaac
     3481 gtcagcaact atgtgaacag cagcgaagtc gtgcccgtga gcagcctcca agagagcagc
     3541 tccaatgagt tcagccccaa caccagcgaa atctgctgca tcatcaggag cagccccggc
     3601 acaaggcaga tgaggagaaa ggaccccgct gtgaagaaca gatgctcccc ccctaaggac
     3661 gtgcctctgg atattatcat gaagaccaac ggaagaacat aagaattcct taaggctagc
     3721 cctaggacgc gtcccgggcc ctatagtgag tcgtattacg tagatccaga catgataaga
     3781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3901 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3961 ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     4021 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4081 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     4141 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4201 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4261 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4321 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4861 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5101 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5161 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5221 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5281 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5341 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5401 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5461 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5521 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5581 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5641 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5701 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5761 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5821 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5881 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5941 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6001 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6061 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6181 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     6241 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6301 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6361 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6421 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6481 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6541 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6601 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     6661 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6721 acgccagtcg accatagcca attcaatatg gcgtatatgg actcatgcca attcaatatg
     6781 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
     6841 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
     6901 gcggacttgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
     6961 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
     7021 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
     7081 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
     7141 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
     7201 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
     7261 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
     7321 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
     7381 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
     7441 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
     7501 ttggcaagta cattactatt ggaaggacgc cagggtacat tggcagtact cccattgacg
     7561 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
     7621 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     7681 ggaggtctat ataagcaatg ctcgtttagg gaac
//
